ArticleRNA (New York, N.Y.)2016
A dual fluorescent reporter for the investigation of methionine mistranslation in live cells.
Article in RNA (New York, N.Y.), 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 20 papers.
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Who cites it
20 citing papers in PubMed.
- Mistranslation from an endogenous tRNA variant in human pan-genome cell lines.Nucleic acids research · 2026Article
- Circadian clock control of ribosome composition promotes rhythmic translation and termination fidelity.Cell reports · 2025Article
- Reactivation of the tRNASer/tRNATyr gene cluster in Arabidopsis thaliana root tips.The Plant cell · 2025Article
- Natural human tRNARNA (New York, N.Y.) · 2025Article
- Glutamine missense suppressor transfer RNAs inhibit polyglutamine aggregation.Molecular therapy. Nucleic acids · 2025Article
- Mechanisms and Delivery of tRNA Therapeutics.Chemical reviews · 2024Review
- Engineered mischarged transfer RNAs for correcting pathogenic missense mutations.Molecular therapy : the journal of the American Society of Gene Therapy · 2024Article
- Article
- Genome-wide screening reveals metabolic regulation of stop-codon readthrough by cyclic AMP.Nucleic acids research · 2023Article
- Genetic Interaction of tRNA-Dependent Mistranslation with Fused in Sarcoma Protein Aggregates.Genes · 2023Article
- Shifting the balance: soluble ADAM10 as a potential treatment for Alzheimer's disease.Frontiers in aging neuroscience · 2023Article
- Transfer RNAs: diversity in form and function.RNA biology · 2021Review
- Modulating Mistranslation Potential of tRNAGenetics · 2019Article
- Pathways to disease from natural variations in human cytoplasmic tRNAs.The Journal of biological chemistry · 2019Review
- Review
- Visualizing translational errors: one cell at a time.Current genetics · 2018Review
- Visualizing tRNA-dependent mistranslation in human cells.RNA biology · 2018Article
- MS-READ: Quantitative measurement of amino acid incorporation.Biochimica et biophysica acta. General subjects · 2017Article
- Determination of tRNA aminoacylation levels by high-throughput sequencing.Nucleic acids research · 2017Article
- Determining the fidelity of tRNA aminoacylation via microarrays.Methods (San Diego, Calif.) · 2017Article
Corrections and comments
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Authors and funding
9 authors.
Funding
Abstract
In mammalian cells under oxidative stress, the methionyl-tRNA synthetase (MetRS) misacylates noncognate tRNAs at frequencies as high as 10% distributed among up to 28 tRNA species. Instead of being detrimental for the cell, misincorporation of methionine residues in the proteome reduces the risk of oxidative damage to proteins, which aids the oxidative stress response. tRNA microarrays have been essential for the detection of the full pattern of misacylated tRNAs, but have limited capacity to investigate the misacylation and mistranslation mechanisms in live cells. Here we develop a dual-fluorescence reporter to specifically measure methionine misincorporation at glutamic acid codons GAA and GAG via tRNA(Glu) mismethionylation in human cells. Our method relies on mutating a specific Met codon in the active site of the fluorescent protein mCherry to a Glu codon that renders mCherry nonfluorescent when translation follows the genetic code. Mistranslation utilizing mismethionylated tRNA(Glu) restores fluorescence in proportion to the amount of misacylated tRNA(Glu). This cellular approach works well for both transient transfection and established stable HEK293 lines. It is rapid, straightforward, and well suited for high-throughput activity analysis under a wide range of physiological conditions. As a proof of concept, we apply this method to characterize the effect of human tRNA(Glu) isodecoders on mistranslation and discuss the implications of our findings.
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